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肾脏病与透析肾移植杂志 ›› 2021, Vol. 30 ›› Issue (1): 7-13.DOI: 10.3969/j.issn.1006298X.2021.01.002

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慢性肾脏病大鼠基于RNA-seq的肠道转录组学分析

  

  • 出版日期:2021-02-28 发布日期:2021-02-27

Intestinal transcriptome analysis of rats with chronic kidney disease based on RNA-Seq technique

  • Online:2021-02-28 Published:2021-02-27

摘要: 目的:分析慢性肾脏病(CKD)大鼠与假手术组大鼠肠道基因表达谱的差异,探讨基因表达差异与CKD肠道变化及并发症的关系。
方法:SpragueDawley大鼠随机分为CKD组与假手术组,CKD组行5/6肾切除术构建CKD模型。28周后收取大鼠血清及空肠组织标本;Masson及天狼猩红染色观察空肠病理变化;每组随机选择3个空肠标本进行RNAseq检测。
结果:CKD组大鼠空肠胶原组成较假手术组差异显著;共筛选出420个差异表达基因(DEGs),CKD大鼠肠道基因表达谱差异显著。GO(gene ontology)及KEGG(Kyoto Encyclopedia of Genes and Genomes)富集分析发现DEGs主要富集到消化系统通路和细胞外基质构建。通过STRING及Cytoscape筛选出Hub基因。
结论:CKD大鼠基因表达差异显著,主要影响消化酶的分泌和细胞外基质的正常结构及功能,可能与CKD患者的胃肠道症状及肠屏障破坏有较大关联。


Abstract: Objective:To analyze the difference of intestinal gene expression profile between rats with chronic kidney disease (CKD) and rats in sham operation group,to explore the relationship between gene expression difference and intestinal changes with complications of CKD.
Methodology:SpragueDawley rats were randomly divided into CKD group, sham operation group. CKD group underwent 5/6 nephrectomy to construct  model of CKD. Rat serum and jejunum tissue samples were collected after 28 weeks. Masson and Wolf scarlet staining were used to observe  pathological changes of jejunum. Three jejunal specimens were randomly selected for RNASeq detection in each group.
Results:CKD model was established successfully. The collagen composition of jejunum in CKD group was significantly different from that in sham operation group. The intestinal gene expression profile of CKD rats was significantly different,and a total of 420 differentially expressed genes (DEGs) were selected. Gene ontology(GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis showed that DEGs were mainly concentrated in digestive system pathways and extracellular matrix construction. Hub genes were screened out through STRING and Cytoscape.
Conclusion:There were significant differences in gene expression in rats with CKD,which mainly affected the secretion of digestive enzymes and normal structure, function of extracellular matrix,which may be significantly associated with gastrointestinal symptoms and intestinal barrier destruction in  CKD.